Shear stress distribution on beam cross-sections under shear loading

In this work, some results obtained by Trabucho and Viaño for the shear stress distribution in beam cross sections using asymptotic expansions of the three-dimensional elasticity equations are compared with those calculated by the classical formulae of the Strength of Materials. We use beams with rectangular and circular cross section to compare the degree of accuracy reached by each method.

A consistent method of analysis of RC shell structures

In this paper some topics related to the design of reinforced concrete (RC) shells are addressed. The influence of the reinforcement layout on the service and ultimate behavior of the shell structure is commented. The well established methodology for dimensioning and verifying RC sections of beam structures is extended. In this way it is possible to treat within a unified procedure the design and verification of RC two dimensional structures, in particular membrane and shell structures. Realistic design situations as multiple steel farnilies and non orthogonal reinforcement layout can be handled. Finally, some examples and applications of the proposed methodology are presented.

Fluorescent quenching-based quantitative detection of specific DNA/RNA using a BODIPY® FL-labeled probe or primer

We have developed a simple method for the quantitative detection of specific DNA or RNA molecules based on the finding that BODIPY® FL fluorescence was quenched by its interaction with a uniquely positioned guanine. This approach makes use of an oligonucleotide probe or primer containing a BODIPY® FL-modified cytosine at its 5′-end. When such a probe was hybridized with a target DNA, its fluorescence was quenched by the guanine in the target, complementary to the modified cytosine, and the quench rate was proportional to the amount of target DNA. This widely applicable technique will be used directly with larger samples or in conjunction with the polymerase chain reaction to quantify small DNA samples.

A novel method for simultaneous high resolution identification of HLA-A, HLA-B, and HLA-Cw alleles.

We describe a novel high resolution DNA based typing approach for HLA class I alleles, which identifies the recombinational motifs present in exons 2 and 3 of the HLA class I genes. Unique identification patterns for 201 known HLA-A, HLA-B, and HLA-Cw alleles were generated by the use of only 40 probes, which were targeted at these common motifs. The unambiguous identification of the alleles was achieved by the development of a new and powerful allelic separation technique that allows isolation of single alleles after amplification. To validate the method, we have used locus-specific primers to amplify exons 2 and 3 of HLA-A, HLA-B, and HLA-Cw loci from 22 heterozygous and 41 homozygous cell lines. After amplification, the allelic fragments from each locus were separated, blotted, and hybridized with the 40 probes. In all cases, the allelic products could be separated and 81 different class I alleles, 33 HLA-A, 30 HLA-B, and 18 HLA-Cw, were identified according to the predicted probe hybridization patterns.

Suppression subtractive hybridization: a method for generating differentially regulated or tissue-specific cDNA probes and libraries.

A new and highly effective method, termed suppression subtractive hybridization (SSH), has been developed for the generation of subtracted cDNA libraries. It is based primarily on a recently described technique called suppression PCR and combines normalization and subtraction in a single procedure. The normalization step equalizes the abundance of cDNAs within the target population and the subtraction step excludes the common sequences between the target and driver populations. In a model system, the SSH technique enriched for rare sequences over 1,000-fold in one round of subtractive hybridization. We demonstrate its usefulness by generating a testis-specific cDNA library and by using the subtracted cDNA mixture as a hybridization probe to identify homologous sequences in a human Y chromosome cosmid library. The human DNA inserts in the isolated cosmids were further confirmed to be expressed in a testis-specific manner. These results suggest that the SSH technique is applicable to many molecular genetic and positional cloning studies for the identification of disease, developmental, tissue-specific, or other differentially expressed genes.

Avaliação e correlação clínica/tomográfica dos biótipos periodontais

O biótipo gengival, definido como a espessura da gengiva no sentido vestíbulo-lingual, desempenha importante papel na homeostasia dos tecidos periodontais, podendo ser considerado um preditor no sucesso em longo prazo das terapias periodontais e periimplantares. Assim sendo, é de suma importância reconhecer as dimensões do tecido gengival e as diferentes formas de qualificá-lo e principalmente quantificá-lo. Apesar de haver descrito na literatura inúmeros métodos para este fim, existem poucos estudos comparando a efetividade de um método em relação a outro. Desta maneira, este estudo buscou avaliar se há concordância entre avaliações clínicas e tomográficas na classificação do biótipo gengival, se existe correlação entre o biótipo gengival e a espessura óssea subjacente, além de descrever um novo método de tomada tomográfica que permita a análise quantitativa do biótipo gengival. Foram avaliados 12 indivíduos os quais realizaram tomografias computadorizadas de feixe cônico como exame imaginológico de diagnóstico ou planejamento pré-cirúrgico. Em cada paciente foram realizados quatro diferentes métodos de avaliação qualitativa da espessura gengival (transparência a sondagem, transgengival, visual através de fotografia e tomográfico), dois métodos de avaliação quantitativa (transgengival e tomográfico) da espessura gengival e avaliação da espessura óssea através da tomografia computadorizada de feixe cônico. Os resultados foram avaliados estatisticamente através do teste KAPPA, Teste t pareado e coeficiente de correlação de Pearson (pM0.05). O novo método de tomada tomográfica descrito neste estudo é eficaz para avaliação do biótipo gengival, havendo grande concordância (86,1% Kappa 0,51) e forte correlação (r=0,824) entre ele e o método transgengival (padrão ouro). A correlação entre a espessura óssea e a espessura gengival foi moderada quando utilizado o método transgengival e tomográfico (r=0,567 e r=0,653 respectivamente).

A detector based on silica fibers for ion beam monitoring in a wide current range

A detector based on doped silica and optical fibers was developed to monitor the profile of particle accelerator beams of intensity ranging from 1 pA to tens of µA. Scintillation light produced in a fiber moving across the beam is measured, giving information on its position, shape and intensity. The detector was tested with a continuous proton beam at the 18 MeV Bern medical cyclotron used for radioisotope production and multi-disciplinary research. For currents from 1 pA to 20 µA, Ce3+ and Sb3+ doped silica fibers were used as sensors. Read out systems based on photodiodes, photomultipliers and solid state photomultipliers were employed. Profiles down to the pA range were measured with this method for the first time. For currents ranging from 1 pA to 3 µA, the integral of the profile was found to be linear with respect to the beam current, which can be measured by this detector with an accuracy of ∼1%. The profile was determined with a spatial resolution of 0.25 mm. For currents ranging from 5 µA to 20 µA, thermal effects affect light yield and transmission, causing distortions of the profile and limitations in monitoring capabilities. For currents higher than ∼1 µA, non doped optical fibers for both producing and transporting scintillation light were also successfully employed.

Exploring Multi-Component Superconducting Compounds by a High-Pressure Method and Ceramic Combinatorial Chemistry

In this short review, we provide some new insights into the material synthesis and characterization of modern multi-component superconducting oxides. Two different approaches such as the high-pressure, high-temperature method and ceramic combinatorial chemistry will be reported with application to several typical examples. First, we highlight the key role of the extreme conditions in the growth of Fe-based superconductors, where a careful control of the composition-structure relation is vital for understanding the microscopic physics. The availability of high-quality LnFeAsO (Ln = lanthanide) single crystals with substitution of O by F, Sm by Th, Fe by Co, and As by P allowed us to measure intrinsic and anisotropic superconducting properties such as Hc2, Jc. Furthermore, we demonstrate that combinatorial ceramic chemistry is an efficient way to search for new superconducting compounds. A single-sample synthesis concept based on multi-element ceramic mixtures can produce a variety of local products. Such a system needs local probe analyses and separation techniques to identify compounds of interest. We present the results obtained from random mixtures of Ca, Sr, Ba, La, Zr, Pb, Tl, Y, Bi, and Cu oxides reacted at different conditions. By adding Zr but removing Tl, Y, and Bi, the bulk state superconductivity got enhanced up to about 122 K.

Analysis of the free vibration of rectangular plates with central cut-outs using the discrete Ritz method

We present an analysis of the free vibration of plates with internal discontinuities due to central cut-outs. A numerical formulation for a basic L-shaped element which is divided into appropriate sub-domains that are dependent upon the location of the cut-out is used as the basic building element. Trial functions formed to satisfy certain boundary conditions are employed to define the transverse deflection of each sub-domain. Mathematical treatments in terms of the continuities in displacement, slope, moment, and higher derivatives between the adjacent sub-domains are enforced at the interconnecting edges. The energy functional results, from the proper assembly of the coupled strain and kinetic energy contributions of each sub-domain, are minimized via the Ritz procedure to extract the vibration frequencies and. mode shapes of the plates. The procedures are demonstrated by considering plates with central cut-outs that are subjected to two types of boundary conditions. (C) 2003 Elsevier Ltd. All rights reserved.

Modeling diffraction and imaging of laser beams by the mode-expansion method

We present a new method of modeling imaging of laser beams in the presence of diffraction. Our method is based on the concept of first orthogonally expanding the resultant diffraction field (that would have otherwise been obtained by the laborious application of the Huygens diffraction principle) and then representing it by an effective multimodal laser beam with different beam parameters. We show not only that the process of obtaining the new beam parameters is straightforward but also that it permits a different interpretation of the diffraction-caused focal shift in laser beams. All of the criteria that we have used to determine the minimum number of higher-order modes needed to accurately represent the diffraction field show that the mode-expansion method is numerically efficient. Finally, the characteristics of the mode-expansion method are such that it allows modeling of a vast array of diffraction problems, regardless of the characteristics of the incident laser beam, the diffracting element, or the observation plane. (C) 2005 Optical Society of America.

Unlocking pathology archives for molecular genetic studies: a reliable method to generate probes for chromogenic and fluorescent in situ hybridization

Chromogenic (CISH) and fluorescent ( FISH) in situ hybridization have emerged as reliable techniques to identify amplifications and chromosomal translocations. CISH provides a spatial distribution of gene copy number changes in tumour tissue and allows a direct correlation between copy number changes and the morphological features of neoplastic cells. However, the limited number of commercially available gene probes has hindered the use of this technique. We have devised a protocol to generate probes for CISH that can be applied to formalin-fixed, paraffin-embedded tissue sections (FFPETS). Bacterial artificial chromosomes ( BACs) containing fragments of human DNA which map to specific genomic regions of interest are amplified with phi 29 polymerase and random primer labelled with biotin. The genomic location of these can be readily confirmed by BAC end pair sequencing and FISH mapping on normal lymphocyte metaphase spreads. To demonstrate the reliability of the probes generated with this protocol, four strategies were employed: (i) probes mapping to cyclin D1 (CCND1) were generated and their performance was compared with that of a commercially available probe for the same gene in a series of 10 FFPETS of breast cancer samples of which five harboured CCND1 amplification; (ii) probes targeting cyclin-dependent kinase 4 were used to validate an amplification identified by microarray-based comparative genomic hybridization (aCGH) in a pleomorphic adenoma; (iii) probes targeting fibroblast growth factor receptor 1 and CCND1 were used to validate amplifications mapping to these regions, as defined by aCGH, in an invasive lobular breast carcinoma with FISH and CISH; and (iv) gene-specific probes for ETV6 and NTRK3 were used to demonstrate the presence of t(12; 15)(p12; q25) translocation in a case of breast secretory carcinoma with dual colour FISH. In summary, this protocol enables the generation of probes mapping to any gene of interest that can be applied to FFPETS, allowing correlation of morphological features with gene copy number.

Optical force field mapping in microdevices

We present a method for characterizing microscopic optical force fields. Two dimensional vector force maps are generated by measuring the optical force applied to a probe particle for a grid of particle positions. The method is used to map Out the force field created by the beam from a lensed fiber inside a liquid filled microdevice. We find transverse gradient forces and axial scattering forces on the order of 2 pN per 10 mW laser power which are constant over a considerable axial range (> 35 mu m). These findings suggest Future useful applications of lensed fibers for particle guiding/sorting. The propulsion of a small particle at a constant velocity of 200 mu m s(-1) is shown.

Rapid quantification of hepatitis B virus DNA by real-time PCR using efficient TaqMan probe and extraction of virus DNA

Aim: To rapidly quantify hepatitis B virus (HBV) DNA by real-time PCR using efficient TaqMan probe and extraction methods of virus DNA. Methods: Three standards were prepared by cloning PCR products which targeted S, C and X region of HBV genome into pGEM-T vector respectively. A pair of primers and matched TaqMan probe were selected by comparing the copy number and the Ct values of HBV serum samples derived from the three different standard curves using certain serum DNA. Then the efficiency of six HBV DNA extraction methods including guanidinium isothiocyanate, proteinase K, NaI, NaOH lysis, alkaline lysis and simple boiling was analyzed in sample A, B and C by real-time PCR. Meanwhile, 8 clinical HBV serum samples were quantified. Results: The copy number of the same HBV serum sample originated from the standard curve of S, C and X regions was 5.7 × 104/ mL, 6.3 × 102/mL and 1.6 × 103/mL respectively. The relative Ct value was 26.6, 31.8 and 29.5 respectively. Therefore, primers and matched probe from S region were chosen for further optimization of six extraction methods. The copy number of HBV serum samples A, B and C was 3.49 × 109/mL, 2.08 × 106/mL and 4.40 × 107/mL respectively, the relative Ct value was 19.9, 30 and 26.2 in the method of NaOH lysis, which was the efficientest among six methods. Simple boiling showed a slightly lower efficiency than NaOH lysis. Guanidinium isothiocyanate, proteinase K and NaI displayed that the copy number of HBV serum sample A, B and C was around 105/ mL, meanwhile the Ct value was about 30. Alkaline failed to quantify the copy number of three HBV serum samples, Standard deviation (SD) and coefficient variation (CV) were very low in all 8 clinical HBV serum samples, showing that quantification of HBV DNA in triplicate was reliable and accurate. Conclusion: Real-time PCR based on optimized primers and TaqMan probe from S region in combination with NaOH lysis is a simple, rapid and accurate method for quantification of HBV serum DNA. © 2006 The WJG Press. All rights reserved.

Specific detection of enterovirus 71 directly from clinical specimens using real-time RT-PCR hybridization probe assay

Enterovirus 71 (EV71) is one of the main causative agents of hand, foot and mouth disease (HFMD) in young children. Infections caused by EV71 could lead to many complications, ranging from brainstem encephalitis to pulmonary oedema, resulting in high mortality. Thus, rapid detection of the virus is required to enable measures to be implemented in preventing widespread transmission. Based on primers and probes targeting at the VP1 region, a real-time reverse-transcriptase polymerase chain reaction (RT-PCR) hybridization probe assay was developed for specific detection of EV71 from clinical specimens. Quantitative analysis showed that the assay was able to detect as low as 5 EV71 viral copies and EV71 was detected from 46 of the 55 clinical specimens obtained from pediatric patients suffering from HFMD during the period from 2000 to 2003 in Singapore. This study showed that the single tube real-time RT-PCR assay developed in this study can be applied as a rapid and sensitive method for specific detection of EV71 directly from clinical specimens. (c) 2005 Elsevier Ltd. All rights reserved.

Evaluation of parameters effective in FRP shear strengthening of RC beams using FE method

Shear strengthening is required when an RC beam is found deficient in shear, or when its shear capacity falls below its flexural capacity after flexural strengthening. A recent technique for the shear strengthening of RC beams is to provide additional FRP web reinforcement, commonly in the form of bonded external FRP strips/sheets. Over the last few years, several experimental studies have been conducted on this new strengthening technique, which has established its effectiveness. While experimental methods of investigation are extremely useful in obtaining information about the composite behaviour of FRP and reinforced concrete, the use of numerical models such as the one presented in this paper helps in developing a good understanding of the behaviour at lower costs. In the study presented in this paper, ANSYS finite element program is used to examine the response of beams strengthened in shear by FRPs. The FE model is calibrated against test results performed at the University of Kentucky. Once validated, the model is used to examine the influence of fibre orientation, compressive strength of concrete, area of tensile and compressive reinforcements, and amount and distance between stirrups on the strength and ductility of FRP strengthened beam.